3 research outputs found
Dynamic DNA Nanotubes: Reversible Switching between Single and Double-Stranded Forms, and Effect of Base Deletions
DNA nanotubes hold great potential as drug delivery vehicles and as programmable templates for the organization of materials and biomolecules. Existing methods for their construction produce assemblies that are entirely double-stranded and rigid, and thus have limited intrinsic dynamic character, or they rely on chemically modified and ligated DNA structures. Here, we report a simple and efficient synthesis of DNA nanotubes from 11 short unmodified strands, and the study of their dynamic behavior by atomic force microscopy and <i>in situ</i> single molecule fluorescence microscopy. This method allows the programmable introduction of DNA structural changes within the repeat units of the tubes. We generate and study fully double-stranded nanotubes, and convert them to nanotubes with one, two and three single-stranded sides, using strand displacement strategies. The nanotubes can be reversibly switched between these forms without compromising their stability and micron-scale lengths. We then site-specifically introduce DNA strands that shorten two sides of the nanotubes, while keeping the length of the third side. The nanotubes undergo bending with increased length mismatch between their sides, until the distortion is significant enough to shorten them, as measured by AFM and single-molecule fluorescence photobleaching experiments. The method presented here produces dynamic and robust nanotubes that can potentially behave as actuators, and allows their site-specific addressability while using a minimal number of component strands
Interaction of Anionic Phenylene Ethynylene Polymers with Lipids: From Membrane Embedding to Liposome Fusion
Here
we report spectroscopic studies on the interaction of negatively
charged, amphiphilic polyphenylene ethynylene (PPE) polymers with
liposomes prepared either from negative, positive or zwitterionic
lipids. Emission spectra of PPEs of 7 and 49 average repeat units
bearing carboxylate terminated side chains showed that the polymer
embeds within positively charged lipids where it exists as free chains.
No interaction was observed between PPEs and negatively charged lipids.
Here the polymer remained aggregated giving rise to broad emission
spectra characteristic of the aggregate species. In zwitterionic lipids,
we observed that the majority of the polymer remained aggregated yet
a small fraction readily embedded within the membrane. Titration experiments
revealed that saturation of zwitterionic lipids with polymer typically
occurred at a polymer repeat unit to lipid mole ratio close to 0.05.
No further membrane embedding was observed above that point. For liposomes
prepared from positively charged lipids, saturation was observed at
a PPE repeat unit to lipid mole ratio of ∼0.1 and liposome
precipitation was observed above this point. FRET studies showed that
precipitation was preceded by lipid mixing and liposome fusion induced
by the PPEs. This behavior was prominent for the longer polymer and
negligible for the shorter polymer at a repeat unit to lipid mole
ratio of 0.05. We postulate that fusion is the consequence of membrane
destabilization whereby the longer polymer gives rise to more extensive
membrane deformation than the shorter polymer
Simple Design for DNA Nanotubes from a Minimal Set of Unmodified Strands: Rapid, Room-Temperature Assembly and Readily Tunable Structure
DNA nanotubes have great potential as nanoscale scaffolds for the organization of materials and the templation of nanowires and as drug delivery vehicles. Current methods for making DNA nanotubes either rely on a tile-based step-growth polymerization mechanism or use a large number of component strands and long annealing times. Step-growth polymerization gives little control over length, is sensitive to stoichiometry, and is slow to generate long products. Here, we present a design strategy for DNA nanotubes that uses an alternative, more controlled growth mechanism, while using just five unmodified component strands and a long enzymatically produced backbone. These tubes form rapidly at room temperature and have numerous, orthogonal sites available for the programmable incorporation of arrays of cargo along their length. As a proof-of-concept, cyanine dyes were organized into two distinct patterns by inclusion into these DNA nanotubes